FIG. 1.

Nucleoporin requirement for nuclear accumulation of activated MAD. (A) S2R+ cells inducibly expressing GFP-MAD and Punt/Tkv (GFP-MAD+R) were treated with the indicated RNAi, and the distribution patterns of GFP-MAD are shown. Nuclei were stained with DAPI. Bar, 10 μM. (B) Two different dsRNA constructs against each nucleoporin hit were tested as described for panel A. The GFP-MAD signals in the nucleus and cytoplasm were quantified, and the ratios were plotted. Data are means ± SEs from ≥3 fields. (C) GFP-MAD+R cells were treated with dsRNAs targeting the 3′ UTR of sec13 or nup93, followed by transfection of HA-Sec13 or HA-Nup93 cDNAs. The distributions of GFP-MAD (green) and the rescue constructs (red) are shown. (D and E) GFP-MAD+R cells were subjected to the indicated RNAi, and the cell extracts were analyzed by immunoblotting with the indicated antibodies.